IBI I-Blue Mini/Midi Kits

The I-Blue MINI Plasmid Kit was designed for rapid isolation of plasmid DNA from 1-7 mL of cultured bacterial cells. I-Blue Lysis Buffer (an optional color indicator) is included with the kit in order to prevent common handling errors, ensuring efficient cell lysis and neutralization. A modified alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA and RNA contaminants. Typical yields are 20-35 µg for high-copy number plasmid or 3-10 µg for low-copy number plasmid from 4 mL of cultured bacterial cells. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed within 15 minutes. The purified plasmid DNA is ready for use in restriction enzyme digestion, ligation, PCR, and sequencing reactions.

Advantages

• Sample: 1-7 mL of cultured bacterial cells
• Yield: Up to 50 µg of pure plasmid DNA
• Format: Plasmid spin column
• Operation Time: Within 15 minutes
• Elution Volume: 30-100 µL
• Kit Storage: Dry at room temperature (15-25°C) for up to 1 year; PD1 and RNase A mixture should be stored at 2-8°C for up to 6 months

Quality Control
The quality of the I-Blue MINI Plasmid Kit is tested on a lot-to-lot basis by isolating plasmid DNA from a 4 mL overnight E. coli (DH5α) culture containing plasmid pBluescript (A600 > 2 U/mL). Following the purification process, a yield of more than 20 µg is obtained and the A260/A280 ratio is between 1.8-2.0. The purified plasmid DNA (1 µg) is used in EcoRI digestion, and analyzed by electrophoresis.

Kit Components

*For IB47171 and IB47172 add provided RNase A to PD1 Buffer then mix by shaking for a few seconds. Check the box on the bottle. PD1 and RNase A mixture should be stored at 2-8°C for up to 6 months. For IB47170 samples, RNase A was already added to PD1.

**If precipitates have formed in PD2 Buffer, warm the buffer in a 37°C water bath, followed by gentle shaking to dissolve.

***Add absolute ethanol (see the bottle label for volume) to Wash Buffer then mix by shaking for a few seconds. Check the box on the bottle. Be sure and close the bottle tightly after each use to avoid ethanol evaporation.